Some properties of adenosine kinase from Ehrlich ascites-tumour cells.
1. Adenosine kinase was measured in dialysed extracts from Ehrlich ascites-tumour cells by a chromatographic procedure. 2. In the absence of added Mg(2+) the K(m) values for ATP and adenosine were 0.22mm and 2.8mum respectively. 3. The maximum velocity of adenosine kinase with free ATP was about three times that with the Mg(2+)-ATP complex. Free Mg(2+) was a non-competitive inhibitor of the reaction. A small amount of added Mg(2+), Mn(2+) or Ca(2+) was required for maximum adenosine kinase activity after cation bound to the enzyme had been released by treatment with p-chloromercuribenzoate and then removed by dialysis. 4. GTP, ITP, deoxy-ATP, deoxy-GTP, CTP, xanthosine triphosphate, UTP and thymidine triphosphate could partially or completely replace ATP as a phosphate donor. 5. The reaction of ATP with adenosine kinase was competitively inhibited by AMP, GMP, IMP, ADP, deoxy-ADP and IDP (K(i) 0.2, 1.1, 5.9, 1.2, 0.5 and 0.78mm respectively). Enzymic activity was markedly affected by the relative concentrations of AMP, ADP and ATP in assay mixtures. 6. The results are discussed in terms of possible mechanisms regulating the rate of adenosine kinase in vivo.
D000818:Animals; D001665:Binding Sites; D002247:Carbon Isotopes; D002286:Carcinoma, Ehrlich Tumor / Q000201:enzymology*; D002384:Catalysis; D002854:Chromatography, Paper; D003956:Dialysis; D007700:Kinetics; D008274:Magnesium; D009711:Nucleotides; D010770:Phosphotransferases / Q000037:antagonists & inhibitors
D002247:Carbon Isotopes; D009711:Nucleotides; D010770:Phosphotransferases; D008274:Magnesium